The Expression Of Calmodulin In Purkinje Cell In Niemann-pick Type C Mice And The Effect Of Tnf-α Sirna On The Degeneration Of Purkinje Cell

Part one The Expression of Calmodulin in Purkinje Cell in Niemann-Pick Type C MiceObjectiveTo investigate the expression of calmodulin in Purkinje cell in Niemann-Pick type C mice.MethodsImmunohitochemistry was adopted to count the number of Purkinje cells in cerebellum, also associate with Western Blot to detect the expressions of three calmodulins, Calbindin-D28k, IP3R1, SERCA2b in the cerebellum of 4, 6, 8 week-old NPC mouse, used wide type(WT) as control, n=6.ResultsCompared with WT, many Purkinje cells were lost and the number was significant decreased, also partial Purkinje cells moved to granular layer. The expressions of Calbindin-D28k and IP3R1 were obviously down-regulated in the 6 and 8 week-old mouse but there was no decrease in the 4 week-old mouse. SERCA2b was also down-regulated in 8 week-old mouse.ConclusionExpressions of the three calmodulins were down-regulated in the degenerative process of Purkinje cell which may lead to the abnormality of calcium's dynamics and be involved in the degeneration of Purkinje cell. Part two The Effect of Lentiviral Vector-mediated TNF-αsiRNA on the Behavior of Two Week-old Niemann-Pick Disease Type C mouseObjective To search the optimal therapic time of TNF-αsiRNA on Niemann-Pick disease type C mouse.MethodsLentiviral-delivered TNF-α-siRNA was injected into the lateral cerebral ventricle of 2 week- old NPC mouse. The mouse was weighed consecutively also coat hanger test and footprint test were adopted to evaluate their motor ability from the 4th to 8th week, using lentivirus-control group and non-injection NPC mouse as negative control.ResultsLoss of weight was significantly delayed in TNF-α-siRNA group; Coat hanger test indicated that the disorder of motor ability was notably improved in TNF-α-siRNA group; Footprint test showed that the step distance of TNF-α-siRNA group was obviously longer than that of the control group.ConclusionUsing TNF-α-siRNA early to treat NPC mouse can improve its behavior better.